This project is designed to enhance the PI's research capability and the skills and interests of our undergraduate and master's students in neuroscience by having them to participate in the PI's ongoing research project in the endogenous cannabinoid system at The University of Texas at Brownsville (a primarily teaching institution with an academically diverse 93% Hispanic community). Cannabinoid, the bioactive component in marijuana and hashish, affects the brain by acting on the brain cannabinoid receptor (CB1R). Natural molecules that activate CB1R are endocannabinoids (eCBs). eCBs are increasingly recognized as critical signaling molecules in many neurobiological phenomena including synaptic plasticity, memory, reward learning, pain, neuroprotection, and addiction; all of which makes understanding the process of eCB-synthesis a matter of great importance. The PI has demonstrated that a key process to initiate the production of eCBs in the hippocampus is a release of calcium ions from the ryanodine receptor (RyR). The discovery was made by artificially-depolarizing neurons. Therefore, the goal of the present proposal is to identify physiological conditions and endogenous molecule(s) that activate RyR in hippocampal neurons for the production of eCBs. Ghrelin, an orexigenic brain peptide hormone, is selected for investigation. Why ghrelin? Because ghrelin and the ghrelin receptor are involved in reward-related memories in which eCBs are also involved. In addition, the PI's preliminary data suggest the activation of the ghrelin receptor stimulates protein kinase A (PKA), which is reported to enhance the function of RyR. Specific Aims of this proposal are: 1) to determine the localization of the ghrelin receptor in the hippocampus, 2) to characterize cytoplasmic calcium dynamics and eCB-synthesis in response to ghrelin, and 3) to determine the extent to which ghrelin regulates hippocampal synaptic plasticity. Experiments are conducted in cultured hippocampal slices with the techniques of patch clamp recording, calcium imaging, immunohistochemistry, siRNA and eGFP transfection. eCBs and their regulation by calcium are topics of great interest to a wide range of biomedical scientists. Thus, the identification of physiological conditions that initiate the production of brain eCBs will provide a critical understanding for potent neuronal signaling systems of eCBs. The project also offers a valuable opportunity for students to obtain laboratory research experiences, which undoubtedly raise their awareness in public health and diseases, and aid them in establishing their professional careers in biomedical research.